Leukocyte counting from a small amount of whole blood using a size-controlled microcavity array
Leukocyte counting from a small amount of whole blood using a size-controlled microcavity array
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DOI：
10.1002/bit.24471
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发表时间：
2012-08-01
影响因子：
3.8
通讯作者：
Matsunaga, Tadashi
中科院分区：
文献类型：
--
作者：
Hosokawa, Masahito;Asami, Marie;Matsunaga, Tadashi
由于缺乏能够从有限的全血中分离所有白细胞的技术，对少量全血中的总白细胞或特定亚群的绝对计数是困难的。在这项研究中，一种配备了尺寸可控的微腔阵列的微流控装置被开发出来，用于高效地从亚微升全血中分离白细胞。微腔阵列可以根据白细胞和其他血细胞的大小和变形能力的差异，将白细胞从全血中分离出来。在排列的微腔中回收的白细胞被连续处理以进行基于图像的免疫表型分析。本装置无需密度梯度离心法或红细胞裂解法等前处理，即可成功回收1亩L全血中90%以上的白细胞。此外，该系统还成功地完成了1MU L全血中人CD_4~+和CD_8~+白细胞的绝对计数，并与常规的流式细胞仪分析结果进行了良好的相关性。我们的微流控设备作为一种护理点白细胞分析系统的工具具有巨大的潜力。生物技术。比昂斯。2012年；109：20172024。(C)2012年威利期刊公司。
Absolute counting of total leukocytes or specific subsets within small amounts of whole blood is difficult due to a lack of techniques that enable separation of all leukocytes from limited amounts of whole blood. In this study, a microfluidic device equipped with a size-controlled microcavity array for highly efficient separation of leukocytes from submicroliters of whole blood was developed. The microcavity array can separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered over 90% of leukocytes in 1?mu L of whole blood without pretreatment such as density gradient centrifugation or erythrocyte lysis. In addition, the proposed system successfully performed absolute enumeration of human CD4+ and CD8+ leukocytes from 1?mu L of whole blood, and the obtained data showed good correlation with conventional flow cytometric analysis. Our microfluidic device has great potential as a tool for a point-of-care leukocyte analysis system. Biotechnol. Bioeng. 2012; 109:20172024. (c) 2012 Wiley Periodicals, Inc.